Improved plaque assay for human coronaviruses 229E and OC43

PeerJ

article

Nicole Bracci¹  Han-Chi Pan²  Caitlin Lehman¹  Kylene Kehn-Hall¹  Shih-Chao Lin³ 

[1] Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University;National Laboratory Animal Center, National Applied Research Laboratories;Bachelor Degree Program in Marine Biotechnology, National Taiwan Ocean University

关键词: Plaque assay;    Coronavirus;    Agarose;    229E;    OC43;    SARS-CoV-2;    COVID-19;   

DOI  :  10.7717/peerj.10639

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

In light of the COVID-19 pandemic, studies that work to understand SARS-CoV-2 are urgently needed. In turn, the less severe human coronaviruses such as HCoV-229E and OC43 are drawing newfound attention. These less severe coronaviruses can be used as a model to facilitate our understanding of the host immune response to coronavirus infection. SARS-CoV-2 must be handled under biosafety level 3 (BSL-3) conditions. Therefore, HCoV-229E and OC43, which can be handled at BSL-2 provide an alternative to SARS-CoV-2 for preclinical screening and designing of antivirals. However, to date, there is no published effective and efficient method to titrate HCoVs other than expensive indirect immunostaining. Here we present an improved approach using an agarose-based conventional plaque assay to titrate HCoV 229E and OC43 with mink lung epithelial cells, Mv1Lu. Our results indicate that titration of HCoV 229E and OC43 with Mv1Lu is consistent and reproducible. The titers produced are also comparable to those produced using human rhabdomyosarcoma (RD) cells. More importantly, Mv1Lu cells display a higher tolerance for cell-cell contact stress, decreased temperature sensitivity, and a faster growth rate. We believe that our improved low-cost plaque assay can serve as an easy tool for researchers conducting HCoV research.

【 授权许可】

CC BY